Molecular and cellular endocrinology · 2024
Development and primary characterization of a human thyroid organoid in vitro model for thyroid metabolism investigation.
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Cristiani Sofia, Bertolini Andrea, Carnicelli Vittoria, Contu Lucia, Vitelli Valentina, Saba Alessandro, Saponaro Federica, Chiellini Grazia, Sabbatini Antonietta Raffaella Maria, Giambelluca Maria Anita, Lenzi Paola, Fornai Francesco, Rossi Leonardo, Materazzi Gabriele, Ambrosini Carlo Enrico, Rutigliano Grazia, Zucchi Riccardo, Bizzarri Ranieri, Ghelardoni Sandra
Аннотация
A 3D thyroid model was developed to address the limitations of 2D cultures and study the effects of compounds like 3-MNT on dehalogenase 1 (IYD) and metabolic activity. Morphology was assessed by TEM, and the expression of tissue-specific genes (TPO, TSHR, PAX8, TTF-1, NIS, IYD, TG) and metabolic features were analyzed using qRT-PCR, immunofluorescence, western blotting, ELISA, and LC-MS/MS, with and without TSH stimulus and 3-MNT treatment. Confocal and TEM analyses confirmed a follicle-like 3D structure. Expression of TPO, NIS, TG, TSH, and PAX markers was significantly higher (p < 0.05) in 3D versus 2D cultures, and ELISA showed increased TG protein production. 3-MNT treatment inhibited IYD activity, indicated by increased MIT and DIT in the media, and significantly altered (p < 0.05) NIS, TG, IYD, TSHR, and TPO expression. These findings suggest 3D thyroid cultures closely replicate tissue traits and functionality, providing a valuable tool for thyroid research.